Fig. 3.
CCT⍺ and ADRP localization in hepatocytes from L-CKO mice. A: Representative confocal photomicrographs of hepatocytes from control (Lap1fl/fl) and L-CKO mice, cultured with or without OA in the medium. The left panels from each genotype show labeling by anti-CCT⍺ Abs, and the right panels show an overlay of labeling with anti-CCT⍺ Abs (red), BODIPY (green), and DAPI (blue). Arrows indicate a nucleus lacking nucleoplasmic CCTα. Scale bars: 10 μm. B: Percentages of nuclei from control and L-CKO hepatocytes, cultured without (-OA) or with (+OA) OA in the media, with diffuse nucleoplasmic CCTα labeling detected by fluorescence microscopy. Columns show mean percentages of nuclei with diffuse nucleoplasmic CCTα localization, black circles indicate data from separate experiments (40–348 nuclei counted for each experiment and condition), and error bars show SEM. ns = not significant, ∗P < 0.05, ∗∗P < 0.01 by one-way ANOVA with Tukey’s multiple comparison test. C: representative 3D image reconstruction from confocal micrographs of a hepatocyte from an L-CKO mouse, cultured with OA in the media and labeled with anti-CCTα Abs (red), BODIPY (green), and DAPI (blue). Scale bar: 10 μm. D: Representative confocal photomicrographs of hepatocytes from an L-CKO mouse; the left panel shows labeling with anti-ADRP Abs, whereas the middle panel shows an overlay of labeling with anti-ADRP Abs (red), BODIPY (green), and DAPI (blue). The right panel shows a zoomed-in view of the area marked with a white square in the middle panel. Arrowhead indicates a nuclear LD lacking ADRP; arrow indicates a cytoplasmic LD with ADRP. Scale bars: 10 μm (left and middle panels) and 1 μm (right panel). ADRP, adipose differentiation-related protein; CCT⍺, CTP:phosphocholine cytidylyltransferase ⍺; OA, oleic acid; LD, lipid droplet; L-CKO mice, Alb-Cre;Lap1fl/fl mice.